Skin test and tick immune status in susceptible and resistant cattle in Brazil

A cutaneous hypersensitivity test (CHT) was used to correlate host resistance to ticks and type of reaction elicited to unfed larval extract-ULE of the cattle tick Boophilus microplus in European and Indian cattle. Twenty calves were separated into four groups of five animals each: naïve or preinfested Indian or European cattle. CHT was induced by intradermal inoculation of 0.1 ml of ULE cattle tick B. microplus (50 μg protein) in the calf ear. Ear thickness was measured using calipers before and 10 min, 1, 2, 6, 18, 24, 48, 72, 96, and 144 h postinoculation (PI). Preinfested European calves showed only an immediate type reaction with maximum response (75% increase in ear thickness) at 10 min PI. On the other hand, preinfested Indian calves presented an immediate response with maximum reaction (70% increase in ear thickness) between 10 min and one hour PI, and a delayed type reaction at 72 h PI (60% increase in ear thickness). These results point out the crucial role of the cellular immune response of cattle in the expression of resistance to cattle tick B. microplus. Skin test might be useful in the ranking of cattle according to the susceptibility/resistance to ticks.

Complete replacement of the mitochondrial genotype in a Bos indicus calf reconstructed by nuclear transfer to a bos taurus oocyte

Due to the exclusively maternal inheritance of mitochondria, mitochondrial genotypes can be coupled to a particular nuclear genotype by continuous mating of founder females and their female offspring to males of the desired nuclear genotype. However, backcrossing is a gradual procedure that, apart from being lengthy, cannot ascertain that genetic and epigenetic changes will modify the original nuclear genotype. Animal cloning by nuclear transfer using host ooplasm carrying polymorphic mitochondrial genomes allows, among other biotechnology applications, the coupling of nuclear and mitochondrial genotypes of diverse origin within a single generation. Previous attempts to use Bos taurus oocytes as hosts to transfer nuclei from unrelated species led to the development to the blastocyst stage but none supported gestation to term. Our aim in this study was to determine whether B. taurus oocytes support development of nuclei from the closely related B. indicus cattle and to examine the fate of their mitochondrial genotypes throughout development. We show that indicus:taurus reconstructed oocytes develop to the blastocyst stage and produce live offspring after transfer to surrogate cows. We also demonstrate that, in reconstructed embryos, donor cell-derived mitochondria undergo a stringent genetic drift during early development leading, in most cases, to a reduction or complete elimination of B. indicus mtDNA. These results demonstrate that cross-subspecies animal cloning is a viable approach both for matching diverse nuclear and cytoplasmic genes to create novel breeds of cattle and for rescuing closely related endangered cattle.

Avaliação do kit TF-Test para o diagnóstico das infecções por parasitas gastrintestinais em ovinos

This study was performed to standardize parasite egg counting in feces of sheep by TF-Test, in addition to compare this test to the Gordon & Whitlock technique (G&W). Twenty-four lambs were artificially infected with Haemonchus contortus throughout 12 weeks. At the end of this time, faecal samples were taken and animals were slaughtered for worm identification and counting. G&W and TF-Test methods were carried out on each fecal sample. Both tests showed Haemonchus eggs in 95.8% of the samples (P>0.05). The correlation coefficients (r) between fecal egg counts (FEC) using G&W × Total Worm Count (TWC) were r=0.52 (not transformed data) and r=0.85 (transformed data); between FEC by TF-Test × TWC were r=0.51 (not transformed data) and r=0.87 (transformed data). Other 100 fecal samples were taken from naturally infected sheep. In these animals, the G&W and TF-Test methods showed 85% and 86% of fecal samples positive for Strongylidea eggs, respectively (P>0.05). Also in those animals, Eimeria oocysts were found in 33% of fecal samples by TF-Test, whereas in the G&W only 12% were positive (P<0.001). For Strongyloides spp., TF-Test showed 15% of positive fecal samples, whereas G&W showed 5% (P<0.05). In conclusion, both methods were efficient to diagnose gastrointestinal nematodes and TF-Test was superior to diagnose oocysts of Eimeria spp. and eggs of Strongyloides spp; conversely, Strongylidea eggs counting using TF-Test was underestimated.

A place preference test in the fish Nile tilapia

The Nile tilapia fish (Oreochromis niloticus) has a high potential to be used as a model in neuroscience studies. In the present study, the preference of the Nile tilapia between a gravel-enriched (GEE), a shelter-enriched (SEE) or a non-enriched (NEE) environment was determined, for developing a place preference model. Nile tilapia had an initial preference for GEE, but after 1 day of observation, the fish stabilized their frequency of visits among compartments. Hence, any stimulus motivating tilapia increase in compartment visiting indicates a positively reinforcing effect. This feature is very useful for the development of new behavioural paradigms for fish in tests using environmental discrimination, such as the conditioning place preference test. © 2006 Elsevier GmbH. All rights reserved.

Ações de vigilância continuada, papel do cão como animal sentinela para toxoplasmose.

Toxoplasmosis is a worldwide zoonosis caused by Toxoplasma gondii and its definitive host is the domestic and wild felids infecting human beings and other warmblooded animals. Dogs are considered a potential risk on the transmission due they can mechanically transmit oocysts to man. In this study, a retrospective analysis of toxoplasmic infection in dog serum samples sent to Serviço de Diagnóstico de Zoonoses/FMVZ-UNESP/Botucatu, SP, in the period of 1998 to 2007 was performed. During this period 1097 serum samples were analyzed by the indirect fluorescent antibody test (IFAT), with 299 (27.25%) positive. The most frequent titer was 16 (42.80%), followed by 64 (37.79%). The results indicate that T.gondii is distributed in the environment showing the role of the dog as sentinel animal to toxoplasmosis to monitor public health actions to the control of this zoonosis.

Dietary fish oil replacement with lard and soybean oil affects triacylglycerol and phospholipid muscle and liver docosahexaenoic acid content but not in the brain and eyes of surubim juveniles Pseudoplatystoma sp

Triplicate groups of juvenile suribim were fed for 183 days one of four different isonitrogenous (47.6% crude protein) and isolipidic (18.7% lipid) diets formulated using three different lipid sources: 100% fish oil (FO, diet 1); 100% pig lard (L, diet 2); 100% soybean oil (SO, diet 3), and FO/L/SO (1:1:1, w/w/w; diet 4). The tissue levels of fatty acids 18:2n - 6 and 18:3n - 3 decreased relative to corresponding dietary fatty acid values. The 20:5n - 3 and 22:6n - 3 composition of muscle and liver neutral lipids were linearly correlated with corresponding dietary fatty acid composition. In contrast, the 22:6n - 3 composition of the brain and eye were similar among treatments. The 22:6n - 3 level was enriched in all tissues, particularly in the neural tissues. Similar results were observed for tissue polar lipids: fatty acids content reflected dietary composition, with the exception of the 22:6n - 3 level, which showed enrichment and no differences between groups. Given these results, the importance of the biochemical functions (transport and/or metabolism) of 22:6n - 3 in the development of the neural system of surubim warrants further investigation. © Springer Science+Business Media B.V. 2008.

Test-day model for milk yield of dairy buffaloes in colombia

The test-day model is the preferred method for genetic evaluations in dairy cattle. For this study, 28372 test-day records of 1220 lactations from 1997 to 2009 were used. The (co)variance components for milk in test-day were estimated using a Uni and multiple-traits repeated animal model with the Restricted Maximum Likelihood method (REML). The Contemporary Group (herd, year, and season of parity) and the age of parity (linear and quadratic) fixed effects, and the additive genetic, permanent environmental, and residual random effects were included in the model. The heritabilities ranged between 0.06 and 0.45 during lactation. The genetic correlations were greater than 0.93. In conclusion, the test-day model is appropriate for the genetic evaluation of dairy buffaloes in Colombia.

Anti-nociceptive effects of Carpolobia lutea G. Don (Polygalaceae) leaf fractions in animal models

Leaves from Carpolobia lutea (Polygalaceae) were screened to establish the antiulcer ethnomedicinal claim and to quantitatively isolate, elucidate the active compounds by semi-preparative HPLC. The anti-nociceptive effects of Carpolobia lutea (CL) G. Don (Polygalaceae) organic leaf extracts were tested in experimental models in mice. The anti-nociceptive mechanism was determined using tail-flick test, acetic acid-induced abdominal constrictions, formalin-induced hind paw licking and the hot plate test. The fractions (ethanol, ethyl acetate, chloroform, n-hexane) and crude ethyl acetate extract of CL (770 mg/kg, i.p.) produced significant inhibitions of both phases of the formalin-induced pain in mice, a reduction in acetic acid-induced writhing as well as and an elevation of the pain threshold in the hot plate test in mice. The inhibitions were greater to those produced by indomethacin (5 mg/kg, i.p.). Ethyl acetate fraction revealed cinnamic and coumaric acids derivatives, which are described for the first time in literature. These cinnamalglucosides polyphenols characterised from CL may in part account for the pharmacological activities. These findings confirm its ethnomedical use in anti-inflammatory pain and in pains from gastric ulcer-associated symptoms. © 2011 Springer Basel AG.

Non-exhaustive test for aerobic capacity determination in running rats

A simple and applicable method for non-exhaustive aerobic evaluation in running rats is described. Wistar rats were submitted to running test at different velocities (10, 15, 20, 25 m/min) with 48 h recovery among them. At each velocity, the rats ran two bouts of 5 min with 2 min of rest between bouts. Blood samples were collected at the end of each bout for lactate determination. For each intensity, delta lactate was calculated and using deltas obtained by four tests, an individual linear interpolation was plotted. The y-intercept of linear interpolation was the null delta lactate equivalent to the critical velocity (CV). To verify the lactate stabilization at CV, the animals were submitted to 25 min of continuous exercise (15, 20, 25 m/min), with blood collection every 5 min. The estimated CV was 16.6±0.7 m/min, with significant linear regressions (R=0.90±0.03). The rats presented maximal lactate steady state (MLSS) at 3.9±0.4 mmol/L, at 20 m/min. The CV was less than MLSS but significantly correlated with this parameter (r=0.78). This non-exhaustive test seems to be valid for the aerobic evaluation of sedentary rats and this protocol underestimates the MLSS in 20%. This test seems to be the interesting method for the evaluation of rats submitted to acute exercise or physical training.

Importância do modelo animal para testar hipóteses sobre a fisiopatologia do binômio diabetes e incontinência urinária feminina

Aims: To discuss the importance of studying animal models to test hypotheses about the mechanisms of urinary continence and pathophysiology of diabetes and urinary incontinence. Source of Data: A literature review was conducted in PubMed and SciELO. The key words used were diabetes, urinary incontinence, urethra, human and rats. Summary of Findings: There is a strong relation between the genesis of urinary incontinence and diabetes mellitus. Due to the similarity of normal distribution of skeletal muscle and urethra anatomy between humans and rats, these animal models have been used in current research about these disorders. Conclusions: The use of rats as an animal model is suitable for experimental studies that test hypotheses about the mechanisms of continence and pathophysiology of the binomial diabetes mellitus and urinary incontinence, thus enabling solutions of great value in clinical practice.

Eficácia do tetra-test como ferramenta de gestão da qualidade do leite cru refrigerado

The effectiveness of a new management tool for screening refrigerated raw milk quality in a dairy industry is provided. Three hundred and seventy-four samples of raw milk from individual producers and 125 samples from bulk milk carriers from the same producers were analyzed. Producers' samples underwent a fast tube reduction test, or Tetra-Test, to estimate total bacteriological charge and their predominant microbiota. Bulk milk samples underwent standard plate count (SPC) as reference method. Tetra-test was effective in screening and assessing the microbiological quality of refrigerated raw milk, since 82.4% of samples with more than 106UFC/mL were readily detected (5 minutes to 2h). The test may be employed as a management tool either in screening or in interventions on the detected problem. In fact, its results provided complementary information on the characteristics of dominant microbiota. Most samples failed in compliance to IN 51 requirements. Further, 211 samples (56.4%) showed high microbial charge and, consequently, poor quality raw milk. Psychrotrophic groups were not yet the main degrading factors of cooled raw milk, albeit present in 76.8% of samples.

Application of micronucleus test and comet assay to evaluate BTEX biodegradation

The BTEX (benzene, toluene, ethylbenzene and xylene) mixture is an environmental pollutant that has a high potential to contaminate water resources, especially groundwater. The bioremediation process by microorganisms has often been used as a tool for removing BTEX from contaminated sites. The application of biological assays is useful in evaluating the efficiency of bioremediation processes, besides identifying the toxicity of the original contaminants. It also allows identifying the effects of possible metabolites formed during the biodegradation process on test organisms. In this study, we evaluated the genotoxic and mutagenic potential of five different BTEX concentrations in rat hepatoma tissue culture (HTC) cells, using comet and micronucleus assays, before and after biodegradation. A mutagenic effect was observed for the highest concentration tested and for its respective non-biodegraded concentration. Genotoxicity was significant for all non-biodegraded concentrations and not significant for the biodegraded ones. According to our results, we can state that BTEX is mutagenic at concentrations close to its water solubility, and genotoxic even at lower concentrations, differing from some described results reported for the mixture components, when tested individually. Our results suggest a synergistic effect for the mixture and that the biodegradation process is a safe and efficient methodology to be applied at BTEX-contaminated sites. © 2012 Elsevier Ltd.

Random regression models using Legendre polynomials or linear splines for test-day milk yield of dairy Gyr (Bos indicus) cattle

Studies investigating the use of random regression models for genetic evaluation of milk production in Zebu cattle are scarce. In this study, 59,744 test-day milk yield records from 7,810 first lactations of purebred dairy Gyr (Bos indicus) and crossbred (dairy Gyr × Holstein) cows were used to compare random regression models in which additive genetic and permanent environmental effects were modeled using orthogonal Legendre polynomials or linear spline functions. Residual variances were modeled considering 1, 5, or 10 classes of days in milk. Five classes fitted the changes in residual variances over the lactation adequately and were used for model comparison. The model that fitted linear spline functions with 6 knots provided the lowest sum of residual variances across lactation. On the other hand, according to the deviance information criterion (DIC) and Bayesian information criterion (BIC), a model using third-order and fourth-order Legendre polynomials for additive genetic and permanent environmental effects, respectively, provided the best fit. However, the high rank correlation (0.998) between this model and that applying third-order Legendre polynomials for additive genetic and permanent environmental effects, indicates that, in practice, the same bulls would be selected by both models. The last model, which is less parameterized, is a parsimonious option for fitting dairy Gyr breed test-day milk yield records. © 2013 American Dairy Science Association.

Ethopharmacological evaluation of the rat exposure test: A prey-predator interaction test

The rat exposure test (RET) is a prey (mouse)-predator (rat) situation that activates brain defensive areas and elicits hormonal and defensive behavior in the mouse. Here, we investigated possible correlations between the spatiotemporal [time spent in protected (home chamber and tunnel) and unprotected (surface) compartments and frequency of entries into the three compartments] and ethological [e.g., duration of protected and unprotected stretched-attend postures (SAP), duration of contact with the rat's compartment] measures (Experiment 1). Secondly, we investigated the effects of systemic treatment with pro- or anti-aversive drugs on the behavior that emerged from the factor analysis (Experiment 2). The effects of chronic (21 days) imipramine and fluoxetine on defensive behavior were also investigated (Experiment 3). Exp. 1 revealed that the time in the protected compartment, protected SAP and rat contacts loaded on factor 1 (defensive behavior), while the total entries and unprotected SAP loaded on factor 2 (locomotor activity). Exp. 2 showed that alprazolam (but not diazepam) selectively changed the defensive factor. Caffeine produced a mild proaversive-like effect, whereas yohimbine only decreased locomotor activity (total entries). Fluoxetine (but not imipramine) produced a weak proaversive-like effect. 5-HT1A/5-HT2 receptor ligands did not change any behavioral measure. In Exp. 3, chronic fluoxetine (but not imipramine) attenuated the defensive behavior factor without changing locomotion. Given that the defensive factor was sensitive to drugs known to attenuate (alprazolam and chronic fluoxetine) and induce (caffeine) panic attack, we suggest the RET as a useful test to assess the effects of panicolytic and panicogenic drugs. © 2012 Elsevier B.V.

Genetic parameters for test-day milk yield, 305-day milk yield, and lactation length in Guzerat cows

Milk production in tropical environments requires the use of crossbreeding systems including breeds well adapted to harsh conditions, but with lower productivities when compared to specialized breeds. Besides the genetic improvement for milk production, lactation lengths also need to be studied for most of these breeds. Accordingly, genetic parameters were estimated for 305-day cumulative milk yield (MY305), test-day milk yield (TDMY), and lactation length (LL) using information from the first lactations of 2816 Guzerat cows selected for milk production in 28 herds in Brazil. Contemporary groups were defined as herd, year and season of the test for TDMY, and as herd, year and season of calving for MY305 and LL. Variance components were estimated with the restricted maximum likelihood method under a multi-trait animal model. Heritabilities estimated for TDMY ranged from 0.16 to 0.24, and were 0.24 and 0.12 for MY305 and LL, respectively. Genetic correlations were high and positive, ranging from 0.51 to 0.99 among TDMY records, from 0.81 to 0.98 between each TDMY and MY305, and from 0.71 to 0.94 between each TDMY and LL. Genetic parameters obtained in this study indicated the possibility of using test-day records for the prediction of breeding values for milk yield in this population of the Guzerat breed. The use of TDMY as selection criteria would result in indirect gains in MY305 and LL. However, the highest response to selection for MY305 would be obtained by direct selection for this trait. © 2012 Elsevier B.V.